• Title/Summary/Keyword: Biological response

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Physico-Chemical Factors on the Growth of Cochlodinium polykrikoides and Nutrient Utilization (Cochlodinium polykrikoides의 성장에 미치는 물리$\cdot$화학적 요인과 영양염 이용)

  • KIM Hyung Chul;LEE Chang Ku;LEE Sam Geun;KIM Hak Gyoon;PARK Chung Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.445-456
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    • 2001
  • In the 1990s, Cochlodinium polykikoides red tide has been annually occurred in the southern coast of Korea and caused the mass damage to the fisheries with a huge amount of economic loss. The present study was done to establish the biological foundation for the elucidation of the mechanism of C. polykikoiaes red tide. The growth response of C. polykikoides to physico-chemical factors such as temperature, salinity, pH, and light intensity were examined using axenic cultures to evaluate the relative importance of these factors on the dynamics of natural populations, It was found that the highest growth conditions were $25^{\circ}C,\;40\%_{\circ}$, pH 7.5, and 7,500 lux, respectively. The tolerable salinity range of growth was relatively wide at an optimum temperature and was reduced to a much narrower range at a sub-optimum temperature. These findings indicate that C. polykikoides is an eurythermal and euryhaline organism. The organism demanded higher light intensity and oceanic pH narrow in its growth. C. polykikoides utilize inorganic nutrients, such as nitrate and ammonium as N, and phosphate as P. The nutritional requirements of C. polykikoides were $40{\mu}M$ for nitrate, $50{\mu}M$ for ammonium, and $5{\mu}M$ for phosphate. The half saturation constant (Ks) for growth was $2.10{\mu}M$ for nitrate, $1.03{\mu}M$ for ammonium, and $0.57{\mu}M$ for phosphate. These values were comparatively smaller than those of other dinoflagellates reported previously. We confirmed that the organism is characterized as an eutrophic species. However, ammonium Ks value is smaller than that of other eutrophic species, This result indicates that C. polykikoides red tide may outbreak in the waters which eutrophication is in progress rather than eutrophicated waters. C. polykikoides preferred ammonium better than nitrate as a nitrogen source when in a growth stage, Therefore, our results indicate that ammonium is more important nutrient on the growth of the organism in comparison with other inorganic nutrients and C. polykikoides red tide is related with the increased ammonium concentration in the coastal waters.

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The development of transgenic maize expressing Actinobacillus pleuropneumoniae ApxIIA gene using Agrobacterium (아그로박테리움을 이용한 Actinobacillus pleuropneumoniae ApxIIA (ApxII toxin) 유전자 발현 옥수수 형질전환체 개발)

  • Kim, Hyun-A;Yoo, Han-Sang;Yang, Moon-Sik;Kwon, Suk-Yoon;Kim, Jin-Seog;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.313-318
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    • 2010
  • To develop edible vaccines for swine, the embryogenic calli (type II) derived from HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vector pMYV611, 613, 616, and V621, 622 and 623 respectively. Six of those vectors carry nptII gene which confers resistance to paromomycin and apxIIA gene producing ApxII toxin which is generated in various serum types of A. pleuropneumoniae as a target gene. The 4,120 callus clones for pMYV611, 5,959 callus clones for pMYV613, 7,581 callus clones for pMYV616, 52,329 callus clones for V621, 48,948 callus clones for V622, and 56,188 callus clones for V623 were inoculated. The frequency of positive response clone was confirmed into range of 2.3% - 4.4% for each vectors by NPTII ELISA kit assay, and the selected callus clones of them were finally 3 callus clones from pMYV611 (0.07%), 4 callus clones from pMYV613 (0.07%), 2 callus clones from pMYV616 (0.03%), 51 callus clones from V621 (0.1%), 72 callus clones from V622 (0.15%), and 102 callus clones from V623 (0.18%) respectively. From the selected callus clones of each binary vector, the integration of the apxIIA gene into maize genome was detected from 2 plants of pMYV613 and 2 plants of V623 by Southern blot analysis.

A study of Brachytherapy for Intraocular Tumor (안구내 악성종양에 대한 저준위 방사선요법에 관한 연구)

  • Ji, Gwang-Su;Yu, Dae-Heon;Lee, Seong-Gu;Kim, Jae-Hyu;Ji, Yeong-Hun
    • The Journal of Korean Society for Radiation Therapy
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    • v.8 no.1
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    • pp.19-27
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    • 1996
  • I. Project Title A Study of Brachytherapy for intraocular tumor II. Objective and Importance of the project The eye enucleation or external-beam radiation therapy that has been commonly used for the treatment of intraocular tumor have demerits of visual loss and in deficiency of effective tumor dose. Recently, brachytherapy using the plaques containing radioisotope-now treatment method that decrease the demerits of the above mentioned treatment methods and increase the treatment effect-is introduced and performed in the countries, Our purpose of this research is to design suitable shape of plaque for the ophthalmic brachytherapy, and to measure absorbed doses of Ir-192 ophthalmic plaque and thereby calculate the exact radiation dose of tumor and it's adjacent normal tissue. III. Scope and Contents of the project In order to brachytherapy for intraocular tumor, 1. to determine the eye model and selected suitable radioisotope 2. to design the suitable shape of plaque 3. to measure transmission factor and dose distribution for custom made plaques 4. to compare with the these data and results of computer dose calculation models IV. Results and Proposal for Applications The result were as followed. 1. Eye model was determined as a 25mm diameter sphere, Ir-192 was considered the most appropriate as radioisotope for brachytherapy, because of the size, half, energy and availability. 2. Considering the biological response with human tissue and protection of exposed dose, we made the plaques with gold, of which size were 15mm, 17mm and 20mm in diameter, and 1.5mm in thickness. 3. Transmission factor of plaques are all 0.71 with TLD and film dosimetry at the surface of plaques and 0.45, 0.49 at 1.5mm distance of surface, respectively. 4. As compared the measured data for the plaque with Ir-192 seeds to results of computer dose calculation model by Gary Luxton et al. and CAP-PLAN (Radiation Treatment Planning System), absorbed doses are within ${\pm}10\%$ and distance deviations are within 0.4mm Maximum error is $-11.3\%$ and 0.8mm, respectively. As a result of it, we can treat the intraocular tumor more effectively by using custom made gold plaque and Ir-192 seeds.

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Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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Biological Effect of Vaccinium uliginosum L. on STZ-induced Diabetes and Lipid Metabolism in Rats (들쭉이 약물에 의해 유도된 당뇨 및 지질대사에 미치는 생리활성 효과)

  • Han, Eun-Kyung;Kwon, Hyuck-Se;Shin, Se-Gye;Choi, Yoon-Hee;Kang, Il-Jun;Chung, Cha-Kwon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.12
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    • pp.1727-1733
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    • 2012
  • This study was conducted to investigate the effects of Vaccinium uliginosum L. (bilberry) on chemically induced diabetes and hypercholesterolemia. Sprague Dawley (SD) rats were divided into six groups, control (CON), bilberry added group (CBB), streptozotocin (STZ)-induced diabetic group (STZ), STZ and bilberry added group (SBB), high fat fed group (HFF) and high fat and bilberry added group (HFB). Diabetes was chemically induced by intravenous injection of 45 mg/kg body weight STZ in citrate buffer (pH 4.5). Serum triglycerides decreased significantly (p<0.05) in the STZ group that was fed bilberry. Additionally, the athrogenic index (AI) decreased significantly (p<0.05) when compared to the STZ group, while the liver triglycerides tended to decrease in the STZ group. HDL-cholesterol also increased significantly in response to bilberry. When compared to the STZ group, steady attenuation of the blood glucose level was observed upon fasting, 15 min, 30 min, 60 min and 120 min after oral glucose administration. The blood glucose level in the bilberry fed group decreased by 24% when compared to STZ group, while the superoxide dismutase (SOD) became significantly higher (p<0.05) in the STZ group when compared to the CON group. Overall, the results of this study suggest that bilberry stimulates lipid metabolism in both the serum and liver and has a positive effect on glucose metabolism in chemically induced diabetic rats.

Immunogenicity, Reactogenicity and Safety of a Combined DTPa-IPV Vaccine Compared with Separate DTPa and IPV Vaccines in Healthy Korean Infants (한국의 건강한 영아를 대상으로 DTPa-IPV 혼합백신을 접종한 경우와 DTPa 백신과 IPV 백신을 각각 투여하였을 경우의 면역원성, 반응원성 및 안전성)

  • Kim, Chang Hwi;Cha, Sung Ho;Shin, Son Moon;Kim, Chun Soo;Choi, Young Youn;Hong, Young Jin;Chey, Myoung Jae;Kim, Kwang Nam;Hur, Jae Kyun;Jo, Dae Sun;Kim, Sung Shin;Lee, Sang Lak;Song, Eun Song;Ramakrishnan, Gunasekaran;Ok, Jin Ju;Van Der Meeren, Olivier;Bock, Hans L.;Kim, Jung Soo
    • Pediatric Infection and Vaccine
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    • v.17 no.2
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    • pp.156-168
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    • 2010
  • Purpose : To compare immunogenicity and reactogenicity of a combined diphtheria-tetanus-acellular pertussis-inactivated poliovirus vaccine (DTPa-IPV, $Infanrix^{TM}$ IPV, GlaxoSmithKline Biologicals) with co-administration of commercially available DTPa and IPV vaccines at separate injection sites (DTPa+IPV). Methods : A total of 458 infants aged 8-12 weeks were randomized to receive three-ose primary vaccination at 2, 4 and 6 months with DTPa-IPV or DTPa+IPV. Blood samples were collected pre and post vaccination for measurement of immune responses. Reactogenicity was assessed following each dose using diary cards. Results : One month post-dose 3, seroprotection rates for anti-diphtheria, anti-tetanus and anti-poliovirus types 1, 2 and 3 were ${\geq}99.5%$ and vaccine response rates to pertussis antigens were at least 98.6% in both DTPa-IPV and DTPa + IPV groups. Non-inferiority between the groups was demonstrated based on pre-defined statistical criteria. Incidences of both local and systemic symptoms were within the same range across both groups with grade 3 symptoms reported following no more than 4.3% of DTPa-IPV doses and 4.5% of DTPa + IPV doses. Two serious adverse events (both pyrexia) after DTPa-IPV administration were considered vaccine-related. Both infants recovered fully. Conclusion : Combined DTPa-IPV vaccine was immunogenic and well tolerated when used as a three-dose primary vaccination course in Korean infants. DTPa-IPV could be incorporated into the Korean vaccination schedule, reducing the number of injections required to complete primary immunization.

In Vivo Immunological Activity in Fermentation with Black Rice Bran (유색미 미강발효물의 면역활성 효과)

  • Kim, Dong-Ju;Ryu, Su-Noh;Han, Sang-Jun;Kim, Hwa-Young;Kim, Jung-Hak;Hong, Seong-Gil
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.273-281
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    • 2011
  • Rice bran is byproducts of the hulling of rice, an important food resource in Korea. Various studies have been reported immune-enhancing effects of rice bran cultured with Lentinus edodes. In particular black rice bran contains anthocyanin, and the effects of antioxidant have been reported. The objective of the this study was to investigate the possible immune-enhancing effects of black rice bran substance extracted from a submerged culture of Lentinus edodes with black rice bran (crude fermentation-polysaccharide, CFP) and products(crude fermentation-polysaccharide-S. cerevisiae CFP-S, crude fermentation-polysaccharide-L. gasseri, CFP-L) which are of secondary fermentation of by using Saccharomyces cerevisiae and Lactobacillus gasseri in the Blab/c male mice. We found that supplementation of CFP, CFP-S and CFP-L enhanced macrophage and splenocyte proliferation compared to the control group(NC) in mice. Also, we measured the concentration of cytokines(IFN-${\gamma}$, TNF-${\alpha}$, IL-6) secreted by activated macrophage and splenocyte. The results of the experiment are that supplementation of CFP and CFP-S increased the macrophage and splenocyte proliferation compared to the control group but supplementation of CFP-L decreased the splenoyte proliferation compared to the control group(without mitogen and treated with LPS). When macrophage and splenocyte were stimulated by CFP and CFP-S supplementation, it was increased IFN-${\gamma}$, TNF-${\alpha}$ and IL-6 concentration compared with the control group. These results suggest that the capacity of CFP and CFP-S seem to act as a potent immune modulator causing augmentation of immune cell activity, and enhance the immue function through regulating cytokine production capacity by activated macrophage and splenocyte in mice.

Chemical Synthesis and Orientation Disruption Bioassay of Sex Pheromone of the Oriental Fruit Moth, Grapholita molests (Busck) (복숭아순나방(Grapholita molests) 성페로몬 합성과 유인력 교란생물검정)

  • Kim, Yong-Gyun;Bae, Sung-Woo;Bae, Soo-Il;Yoon, Hyang-Mi;Hong, Yong-Pyo
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.309-316
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    • 2006
  • Three sex pheromone components (cis-8-dodecenyl acetate (Z12Ac), trans-8-dodecenyl acetate (E12Ac), cis-8-dodecenol (Z12OH)) of the Oriental fruit moth, Grapholita molesta, were chemically synthesized. Especially to increase the composition of cis-stereoisomer, a triple bond intermediate was hydrogenated at $-20^{\circ}C$ with catalytic $Pd/BaSO_{4}$. The resulting product consisted of the acetates with a stereoisomer ratio in 92:8 (Z:E). The biological activity of the synthesized pheromone compounds was analyzed both in male responses and orientation disruption. The indoor pheromone effect was determined by male flight behavior showing wing movement in response to lure. Different mixtures of the synthetic pheromone components were prepared by mixing acetate and alcohol components in 100:0, 99:1, and 90:10 (g/g) and tested with a comparison of a standard commercial pheromone lure. The highest pheromone effect was observed in only acetate mixture (100:0) and the effect was reduced with the addition of the alcohol component. This indoor pheromone effect could be observed in field monitoring trial, in which 100:0 mixture showed the highest trap catches. Orientation disruption assay was conducted indoor by using a cage, in which the center had a commercial lure on sticky plate and the four candidates were placed at 6 cm away from the central lure on each of four directions. Test males were released to the arena during overnight (12 h) and then the caught males on the sticky plate were counted. The synthesized pheromone as well as the commercial pheromone showed 100% orientation disruption. However, the orientation disruption effect was reduced with decrease in the number of the surrounding disrupting pheromone baits. These results clearly suggest that the synthesized sex pheromone of G. molesta is biologically active and can be used for field mating disruption.